We opted to use the reducing SDS run buffer in conjunction with the Optiblot WB kits. The main driver for this was cost and the ease of use/preparation. The buffers are diluted prior to use and work very well for our applications in standard WB techniques. When used in conjunction with Optiblot gels the results are excellent with clearly defined bands. We're very pleased with the Optiblot range as it has significantly increased our throughput.
We used a typical protocol:
Protein was extracted using RIPA buffer extraction, followed by treatment at 37 degrees Celsius for 30 minutes in Optiblot LDS Sample Buffer (4X). 100ug was loaded per well and run for 45 minutes at 180V. Following transfer and staining we see nice clearly defined bands for our samples.